Crystal form of orexin receptor antagonist compound, and preparation method and application thereof

ABSTRACT

Disclosed is a preparation method of an orexin receptor antagonist compound 5-3. Crystalline forms I-IV of an orexin receptor antagonist compound 5-3 are provided. Also provided are processes of preparing crystalline forms of the orexin receptor antagonist compound 5-3, as well as methods of using the crystalline forms I-IV for treating an orexin-related disease.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a Divisional of U.S. patent application Ser. No.15/752,290, filed Feb. 13, 2018, which is a Section 371 of InternationalApplication No. PCT/CN2016/094558, filed Aug. 11, 2016, which waspublished in the Chinese language on Feb. 23, 2017, under InternationalPublication No. WO 2017/028732 A1, which claims priority under 35 U.S.C.§ 119(b) to Chinese Application No. 201510501331.8, filed Aug. 14, 2015,the disclosures of which are incorporated herein by reference in theirentirety.

TECHNICAL FIELD

The present invention relates to crystalline form I, crystalline formII, crystalline form III and crystalline form IV of the compound 5-3 asorexin receptor antagonist and its preparation method, and itsapplication in the manufacture of medicaments for the treatment oforexin-related diseases.

BACKGROUND

Orexin (or orexins) includes two neuropeptides produced by thehypothalamus: the orexin A (OX-A) (a peptide with 33 amino acids) andthe orexin B (OX-B) (a peptide with 28 amino acids) (Sakurai T., et al.,Cell, 1998, 92, 573-585). It is found that orexin can stimulate foodconsumption in rats, that is to say, in the center feedback mechanism ofregulation of feeding behavior, the peptide has a physiological role asa medium (Sakurai T. et al., Cell, 1998, 92, 573-585). Orexin canregulate sleep and insomnia status, thereby potentially providing a newmethod for treatment of sleep in patients with insomnia or paroxysmal(Chemelli R. M. et al., Cell, 1999, 98, 437-451). Orexin also play arole in awakening, motivation, learning and memory (Harris, et al.,Trends Neuroscl., 2006, 29 (10), 571-577). Two orexin receptors havebeen cloned and characterized in mammals, which belong to the Gprotein-coupled receptor superfamily (Sakurai T. et al., Cell, 1998, 92,573-585): the orexin-1 receptor (OX or OXIR) is selective for OX-A, andorexin-2 receptor (OX2 or OX2R) is capable of binding OX-A and OX-B. Itis believed that the physiological role of orexin is assumed to beexpressed with either or both of OXI receptor and OX2 (two subtypes oforexin receptor).

Orexin receptors can be found in brain of warm-blooded animals, and areinvolved in many diseases, e.g., depression; anxiety; addiction;obsessive compulsory disorder; affective neurosis; depressive neurosis;anxiety neurosis; dysthymic disorder; behavioral disorders; mooddisorders; sexual dysfunction; psychosexual dysfunction; genderdisorder; schizophrenia: manic depression; delirium; dementia; severemental retardation and movement disorders, such as Huntington's diseaseand Tourette syndrome; eating disorders such as anorexia, bulimia,cachexia, and obesity; addictive feeding behavior; binge eating crashfeeding behavior; cardiovascular disease; diabetes; appetite or tastedisorders; emesis, vomiting, nausea; asthma: cancer, Parkinson'sdisease; Cushing's syndrome/disease; basophil adenoma; prolactinoma;hyperprolactinemia; pituitary tumor or adenoma; hypothalamic disease;inflammatory bowel disease; gastric dysfunction; gastric ulcer; obesitygenital degradation; pituitary disorders; pituitary gland disorders;pituitary hypogonadism; pituitary hyperactivity; hypothalamichypogonadism; Kallmann's comprehensive disease (anosmia, hyposmia);functional or psychogenic amenorrhea; hypopituitarism; hypothalamichypothyroidism; hypothalamus-adrenal dysfunction; suddenhyperprolactinemia; hypothalamic disease growth hormone deficiency;sudden lack of growth; dwarfism; gigantism; acromegaly; disturbedbiological and circadian rhythms; sleep disorders associated withdiseases such as neurological disorders, neuropathic pain and restlessleg syndrome; heart and lung diseases, acute and congestive heartfailure; hypotension; hypertension; urinary retention; osteoporosis;angina; acute myocardial infarction; ischemic or hemorrhagic stroke;subarachnoid hemorrhage; ulcers; allergic reaction; benign prostatichypertrophy; chronic renal failure; kidney disease; impaired glucosetolerance; migraine; hyperalgesia; pain; enhanced or exaggeratedsensitivity to pain such as hyperalgesia, causalgia and allodynia; acutepain; burning pain; atypical facial pain; neuropathic pain; back pain;complex regional pain syndrome I and II; arthritic pain; sports injurypain; and infections (such as HIV) related pain, post-chemotherapy pain;post-stroke pain; post-operative pain; neuralgia; vomiting, nausea,vomiting; visceral pain related disorders, such as irritable bowelsyndrome and angina; migraine; urinary bladder incontinence, forexample, urge incontinence; tolerance to narcotics or anesthetics; sleepdisorders; sleep apnea; narcolepsy; insomnia; parasomnia; jet lagsyndrome; and neurodegenerative disorders, including diseaseclassification entities such asdisinhibition-dementia-Parkinson's-muscular atrophy syndrome; epilepsy:seizure disorders and other general orexin system dysfunction relateddiseases.

Preparation of intermediate of ester compounds through hydrogenationreduction of alkene has been reported in the previous literatures.WO2009153178, WO2012036997, WO2012058127 have reported hydrogenationreduction using palladium carbon and palladium hydroxide. However, noneof these references studied the effect of hydrogenation conditions oncis- and trans-products.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a preparation methodof compound 5-3, which includes the following steps:

the reaction pressure is normal pressure to 10 Mpa;

optionally, the reaction temperature is 0° C. to room temperature;

the reaction solvent is selected from the polar organic solvent.

In one embodiment of the present invention, a:b>3:1.

In one embodiment of the present invention, the reaction pressure is 1Mpa to 5 Mpa.

In one embodiment of the present invention, optionally, the reactiontemperature is 5 to 10° C.

In one embodiment of the present invention, the above mentioned polarorganic solvent is selected from ethyl acetate, tetrahydrofuran,methanol, ethanol, isopropanol, or dioxane.

The object of the present invention is to provide a preparation methodof compound 5-3, which includes the following steps:

wherein,

the separation agent is selected from D-tartaric acid;

the reaction solvent is selected from ethyl acetate, isopropyl acetate,ethanol, methanol, tetrahydrofuran, dichloromethane or water;

the recrystallization solvent is selected from water, or ethanol;

the base is selected from sodium carbonate, potassium carbonate, sodiumhydroxide, or potassium hydroxide.

In one embodiment of the present invention, the preparation method ofcompound 5-3 includes the following steps:

wherein the condensation agent is selected from T₃P, CDI, EDCI, HOBt orHATU.

The object of the present invention is to provide crystal form I ofcompound 5-3, whose structure is shown in FIG. 1.

In one embodiment of the present invention, the XRPD pattern analysis ofcrystal form I of compound 5-3 is as follows.

diffraction angle 2θ relative intensity (%) 8.611 100.0 10.330 70.612.284 13.4 13.703 23.9 14.057 5.6 14.351 4.8 14.848 9.2 15.576 7.516.602 31.6 17.250 71.3 17.902 90.7 18.277 42.4 19.149 6.5 20.625 21.721.550 42.7 22.124 2.3 23.307 33.4 23.953 2.6 24.370 32.3 26.501 29.426.915 8.7 27.663 11.0 27.939 15.5 29.499 2.6 30.604 2.2 31.727 13.332.969 2.6 33.378 2.8 33.777 3.0 34.825 1.6 36.399 5.3 37.366 4.3 39.3573.3

The present invention also provides a method for preparing crystal formI, which comprises that adding compound 5-3 into a polar organicsolvent, heating to a temperature from 40° C. to reflux temperature fordissolution, and then slowly adding a weak polar or non-polar organicsolvent dropwise, and after the addition is completed, cooling to atemperature of 0-20° C. in 1-10 hours, thereby forming the crystal.

In one embodiment of the present invention, the above polar organicsolvent is selected from C₁₋₆ alkyl alcohol, C₂₋₆ ester, acetonitrileand/or dichloromethane, or a mixture thereof.

In one embodiment of the present invention, the above C₁₋₆ alkyl alcoholis selected from methanol, ethanol, isopropanol, and/or n-butanol.

In one embodiment of the present invention, the above C₂₋₆ ester isselected from ethyl formate, ethyl acetate, isopropyl acetate, isobutylacetate, and/or n-butyl acetate.

In one embodiment of the invention, the above weak polar or non-polarorganic solvent is selected from C₅₋₁₀ alkane or cycloalkane, C₄₋₁₀ether or cyclic ether, petroleum ether, and/or benzene optionallysubstituted by 1 to 3 methyl or ethyl or halogen atoms.

In some embodiments of the present invention, the above C₅₋₁₀ alkane orcycloalkane is selected from pentane, n-hexane, cyclohexane, n-heptaneand/or isooctane.

In one embodiment of the invention, the above C₄₋₁₀ ether or cyclicether is selected from ethyl ether, methyl tert-butyl ether, n-propylether, n-butyl ether, ethylene glycol dimethyl ether, tetrahydrofuran,dimethyl tetrahydrofuran and/or dioxane.

In one embodiment of the present invention, the above benzene optionallysubstituted by 1 to 3 of methyl or ethyl or halogen atoms is selectedfrom methylbenzene, xylene, or chlorobenzene.

In one embodiment of the present invention, the amount (weight/volume)ratio of compound 5-3 to the polar organic solvent is 1:1-10.

In one embodiment of the present invention, the amount (weight/volume)ratio of compound 5-3 to the polar organic solvent is specifically1:2-5.

In one embodiment of the present invention, the amount (weight/volume)ratio of compound 5-3 to the weak polar or non-polar organic solvent is1:1-15.

In one embodiment of the present invention, the amount (weight/volume)ratio of compound 5-3 to the weak polar or non-polar organic solvent isspecifically 1: 2-7.

The present invention also provides crystal form II of compound 5-3,whose structure is shown in FIG. 2.

In one embodiment of the present invention, the XRPD pattern analysis ofcrystal form II is as follows.

diffraction angle 2θ relative intensity (%) 10.428 60.0 11.968 100.013.542 17.8 14.238 3.4 14.767 12.1 15.851 73.9 16.818 72.0 18.003 9.219.087 2.1 19.755 47.6 20.900 25.7 22.652 2.9 23.858 49.2 24.844 23.225.533 8.2 26.083 11.0 27.133 3.1 27.506 17.7 28.316 3.0 29.557 2.230.740 4.2 31.846 5.1 32.550 2.0 33.775 3.1 34.682 2.2 36.422 3.4 36.9533.4 38.569 3.0

The present invention also provides a method for preparing crystal formII, which comprises that adding crystal form I or crystal form III orcrystal form IV of compound 5-3, or a mixed crystal form or any otherform composed of the three crystal forms in any ratio into an organicsolvent or a mixed solvent of organic solvent and water, and stirringfor 5-120 hours at 20-40° C.

In one embodiment of the present invention, the above organic solvent isselected from C₁₋₆ alkyl alcohol, C₅₋₁₀ alkane or cycloalkane, C₄₋₁₀ether or cyclic ether, C₃₋₇ ketone, C₂₋₆ ester, acetonitrile and/orbenzene optionally substituted by 1-3 methyl or ethyl or halogen atoms.

In one embodiment of the present invention, the C₁₋₆ alkyl alcohol isselected from methanol, ethanol, isopropanol, and/or n-butanol.

In one embodiment of the present invention, the C₅₋₁₀ alkane orcycloalkane is selected from pentane, n-hexane, n-heptane and/orcyclohexane.

In one embodiment of the present invention, the above C₄₋₁₀ ether orcyclic ether is selected from methyl tert-butyl ether, tetrahydrofuran,dimethyl tetrahydrofuran and/or 1,4-dioxane.

In one embodiment of the present invention, the C₃₋₇ ketone is selectedfrom acetone, methyl isobutyl ketone, and/or methyl ethyl ketone.

In one embodiment of the present invention, the C₂₋₆ ester is selectedfrom ethyl acetate, and/or isopropyl acetate.

In one embodiment of the present invention, the benzene optionallysubstituted by 1-3 of methyl or ethyl or halogen atoms is selected frommethylbenzene, xylene, or chlorobenzene.

In one embodiment of the present invention, the mixed solvent of organicsolvent and water is selected from methanol/water, ethanol/water,isopropanol/water, acetonitrile/water, acetone/water,tetrahydrofuran/water or 1,4-dioxane/water.

In one embodiment of the present invention, the volume ratio of organicsolvent to water is 0.1-20:1.

In some embodiments of the present invention, the organic solvent isselected from isopropanol, acetonitrile, methyl isobutyl ketone, ethylacetate, isopropyl acetate, dimethyl tetrahydrofuran, toluene, and/ormethanol.

In one embodiment of the present invention, the volume ratio of organicsolvent to water is specifically 0.5-5:1.

The present invention also provides crystal form III of compound 5-3,whose structure is shown in FIG. 3.

In one embodiment of the present invention, the XRPD pattern analysis ofcrystal form III is as follows.

diffraction angle 2θ relative intensity (%) 8.539 57.2 10.296 65.112.283 8.3 14.023 6.0 14.319 4.0 15.879 1.6 17.230 100.0 17.804 11.518.005 22.4 19.269 1.3 20.591 18.6 21.514 18.5 23.092 2.6 23.322 0.924.319 28.5 26.426 7.1 30.568 1.0 31.686 4.6 33.739 0.9 34.768 0.936.366 5.1 37.326 0.5 39.285 1.3

The present invention also provides a method for preparing crystal formIII, which comprises that adding crystal form I of compound 5-3 or anyother form of compound 5-3 except crystal form II into a polar organicsolvent, dissolving at room temperature or heating to a temperature of40° C. to reflux temperature for dissolution, and then slowly adding aweak polar or non-polar organic solvent dropwise, and after the additionis completed, forming the crystal at room temperature or by cooling to20-30° C. in 1-120 hours.

In one embodiment of the present invention, the above organic solvent isselected from C₁₋₆ alkyl alcohol, C₅₋₁₀ alkane or cycloalkane, C₄₋₁₀ether or cyclic ether, C₃₋₇ ketone, C₂₋₆ ester, acetonitrile and/orbenzene optionally substituted by 1-3 methyl or ethyl or halogen atoms.

In one embodiment of the present invention, the above C₁₋₆ alkyl alcoholis selected from methanol, ethanol, isopropanol, and/or n-butanol.

In one embodiment of the present invention, the C₅₋₁₀ alkane orcycloalkane is selected from pentane, n-hexane, n-heptane and/orcyclohexane.

In one embodiment of the present invention, the C₄₋₁₀ ether or cyclicether is selected from methyl tert-butyl ether, tetrahydrofuran,dimethyl tetrahydrofuran and/or 1,4-dioxane.

In one embodiment of the present invention, the above C₃₋₇ ketone isselected from acetone, methyl isobutyl ketone, and/or methyl ethylketone.

In one embodiment of the present invention, the above C₂₋₄ ester isselected from ethyl acetate, and/or isopropyl acetate.

In one embodiment of the present invention, the benzene optionallysubstituted by 1-3 methyl or ethyl or halogen atoms is selected frommethylbenzene, xylene, or chlorobenzene.

In one embodiment of the present invention, the mixed solvent of organicsolvent and water is selected from methanol/water, ethanol/water,isopropanol/water, acetonitrile/water, acetone/water,tetrahydrofuran/water or 1,4-dioxane/water.

In one embodiment of the present invention, the volume ratio of organicsolvent to water is 0.1-20:1.

In some embodiments of the present invention, the organic solvent isselected from isopropanol, acetonitrile, methyl isobutyl ketone, ethylacetate, isopropyl acetate, dimethyl tetrahydrofuran, toluene, and/ormethanol.

In one embodiment of the present invention, the volume ratio of organicsolvent to water is specifically 0.5-5:1.

The present invention also provides crystal form IV of compound 5-3, thestructure of which is shown in FIG. 3.

In one embodiment of the present invention, the XRPD pattern analysis ofcrystal form IV is as follows.

diffraction angle 2θ relative intensity (%) 8.653 100.0 10.408 94.712.106 45.5 12.362 15.5 13.723 14.5 14.152 6.1 14.414 7.3 14.925 10.615.988 18.9 16.682 20.1 16.937 33.6 17.311 79.1 17.982 61.7 18.338 29.219.245 5.9 19.856 29.2 20.721 24.1 21.020 8.2 21.611 34.8 23.385 22.123.997 44.0 24.430 30.5 24.965 15.2 26.225 9.3 26.562 23.9 26.977 6.827.647 16.3 27.983 10.9 29.659 3.1 30.842 3.0 31.788 12.4 33.031 2.733.426 2.2 33.858 2.8 34.884 2.3 36.499 4.8 37.427 2.9

The present invention also provides a method for preparing crystal formIV, which comprises that adding crystal form I of compound 5-3 or anyother form of compound 5-3 except crystal form II into a polar organicsolvent, dissolving at room temperature or heating to a temperature of40° C. to reflux temperature for dissolution, then slowly adding a weakpolar or non-polar organic solvent dropwise, and after the addition iscompleted, forming the crystal at room temperature or by cooling to20-30° C. in 1-120 hours.

In one embodiment of the present invention, the above organic solvent isselected from C₁₋₆ alkyl alcohol, C₅₋₁₀ alkane or cycloalkane, C₄₋₁₀ether or cyclic ether, C₃₋₇ ketone, C₂₋₆ ester, acetonitrile and/orbenzene optionally substituted by 1-3 methyl or ethyl or halogen atoms.

In one embodiment of the present invention, the above C₁₋₆ alkyl alcoholis selected from methanol, ethanol, isopropanol, and/or n-butanol.

In one embodiment of the present invention, the C₅₋₁₀ alkane orcycloalkane is selected from pentane, n-hexane, n-heptane and/orcyclohexane.

In one embodiment of the present invention, the above C₄₋₁₀ ether orcyclic ether is selected from methyl tert-butyl ether, tetrahydrofuran,dimethyl tetrahydrofuran and/or 1,4-dioxane.

In one embodiment of the present invention, the above C₃₋₇ ketone isselected from acetone, methyl isobutyl ketone, and/or methyl ethylketone.

In one embodiment of the present invention, the above C₂₋₆ ester isselected from ethyl acetate, and/or isopropyl acetate.

In one embodiment of the present invention, the benzene optionallysubstituted by 1-3 methyl or ethyl or halogen atoms is selected frommethylbenzene, xylene, or chlorobenzene.

In one embodiment of the present invention, the mixed solvent of organicsolvent and water is selected from methanol/water, ethanol/water,isopropanol/water, acetonitrile/water, acetone/water,tetrahydrofuran/water or 1,4-dioxane/water.

In one embodiment of the present invention, the volume ratio of organicsolvent to water is 0.1-20:1.

In some embodiments of the present invention, the organic solvent isspecifically selected from isopropanol, acetonitrile, methyl isobutylketone, ethyl acetate, isopropyl acetate, dimethyl tetrahydrofuran,toluene, and/or methanol.

In one embodiment of the present invention, the volume ratio of organicsolvent to water is specifically 0.5-5:1.

Another object of the present invention is to provide a use of crystalform I or crystal form II or crystal form III or crystal form IV ofcompound 5-3 as orexin receptor antagonist in the manufacture of amedicament for the treatment or prevention of neurological andpsychiatric disorders or orexin-related diseases.

In one embodiment of the present invention, the above mentioned use ofcrystal form I or crystal form II or crystal form III or crystal form IVof compound 5-3 as orexin receptor antagonist in the manufacture of amedicament for the treatment or prevention of neurological andpsychiatric disorders or orexin-related diseases, wherein theneurological and psychiatric disorders or orexin-related diseasesinclude insomnia, chronic obstructive pulmonary disease, obstructivesleep apnea, hypersomnia, anxiety, obsessive-compulsive disorder, panicattack, nicotine addiction, or binge eating disorder.

Definition and Description

Unless otherwise stated, the following terms and phrases used herein areintended to have the following meanings. A particular term or phraseshould not be considered as indefinite or unclear unless it isspecifically defined and should be understood in accordance with theordinary meaning.

When a trade name appears in this article, it is intended to refer toits corresponding product or its active ingredient.

C₁₋₁₂ is selected from C₁, C₂, C₃, C₄, C₅, C₆, C₇, C₈, C₉, C₁₀, C₁₁ andC₁₂; C₃₋₁₂ is selected from C₃, C₄, C₅, C₆, C₇, C₈, C₉, C₁₀, C₁₁ andC₁₂.

The term “substituted” refers to any one or more hydrogen atom onparticular atoms is (are) substituted by a substituent, includingdeuterium and variants of hydrogen, as long as the particular atom is ofnormal valence and the compound is stable after substitution. When asubstituent is ketone group (i.e., ═O), it means that two hydrogen atomsare replaced. Ketone group substitution does not occur on the aromaticgroup. The term “optionally substituted” means that it may besubstituted, or may be unsubstituted. Unless otherwise specified, thetype and number of substituents may be arbitrary as long as it ischemically achievable.

When any variable (e.g., R) occurs more than once in the composition orstructure of the compound, its definition in each case are independent.Thus, for example, if a group is substituted with 0-2 of R, then thegroup may optionally be substituted with up to two R, and R in each casecan be independent. In addition, the combination of substituents and/orthe variants thereof is allowed only if such combination results in astable compound.

Unless otherwise specified, the term “halogen element” or “halogen” byitself or as a part of another substituent denotes fluorine, chlorine,bromine or iodine atom. Furthermore, the term “haloalkyl” means toinclude monohaloalkyl and polyhaloalkyl. For example, the term “halo(C1-C4) alkyl” is intended to include, but not limited to,trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl and 3-bromopropyland the like.

The term “halogen” or “halo” refers to fluorine, chlorine, bromine andiodine.

Unless otherwise specified, “ring” represents a substituted orunsubstituted cycloalkyl, heterocycloalkyl, cycloalkenyl,heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl, orheteroaryl. The ring includes a monocyclic ring, bicyclic ring, spiroring, fused ring or bridge ring. The number of ring atoms is usuallydefined as the member number in a ring, for example, “5 to 7-memberedring” means 5 to 7 atoms are arranged in a cycle. Unless otherwisespecified, the ring optionally contains 1 to 3 hetero atoms. Therefore,“5 to 7-membered ring” includes for example, phenyl pyridine andpiperidinyl; on the other hand, the term “5 to 7-memberedheterocycloalkyl ring” include pyridyl and piperidyl, but excludingphenyl. The term “ring” also includes ring systems containing at leastone ring, wherein each “ring” independently meets the above definition.

Unless otherwise specified, the term “hydrocarbyl” or the specificconcept (such as alkyl, alkenyl, alkynyl, phenyl, etc.) by itself or asa part of another substituent represents a straight, branched or cyclichydrocarbon radical or combinations thereof, may be fully saturated,mono- or poly-unsaturated, may be mono-, di- or poly-substituted, can bemonovalent (eg methyl), divalent (eg methylene) or polyvalent (egmethine), and may include divalent or multivalent radicals, and hascertain number of carbon atoms (for example, C1-C10 indicates 1 to 10carbons). “Hydrocarbyl” includes but not limited to aliphatichydrocarbon groups and aromatic hydrocarbon groups. The aliphatichydrocarbon groups include linear and cyclic groups, and include but arenot limited to alkyl group, alkenyl group, and alkynyl group; thearomatic hydrocarbon groups include but are not limited to 6-12 memberedaromatic hydrocarbon groups, such as benzene, naphthalene and the like.In some embodiments, the term “alkyl” represents linear, branchedradicals or the combinations thereof, which may be fully saturated,mono- or poly-unsaturated and can include di- or multivalent radicals.Examples of saturated hydrocarbon radicals include, but are not limitedto, methyl, ethyl, n-propyl, isopropyl, n-butyl, tertiary butyl,isobutyl, sec-butyl, isobutyl, cyclohexyl, (cyclohexyl) methyl,cyclopropylmethyl, and homologs or isomers of n-pentyl, n-hexyl,n-heptyl, n-octyl radicals. Unsaturated alkyl groups have one or moredouble or triple bonds, examples of which include but are not limitedto, ethenyl, 2-propenyl, butenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and higher homologs and isomers.

Unless otherwise specified, the term “heterohydrocarbyl”, or thespecific concepts thereof (e.g. heteroalkyl, heteroalkenyl,heteroalkynyl, heteroaryl, etc.) by itself or in combination with otherterms indicates a stable straight chain, branched chain or cyclichydrocarbon radical, or the combinations thereof and comprises a certainnumber of carbon atoms and at least one hetero atoms. In someembodiments, the term “heteroalkyl” by itself or in combination withother terms indicates a stable straight chain, branched chain, or thecombinations thereof and comprises a certain number of carbon atoms andat least one hetero atoms. In one exemplary embodiment, a heteroatom isselected from B, O, N and S, wherein the nitrogen and sulfur atoms areoptionally oxidized and the nitrogen atom is optionally quaternized.Heteroatoms B, O, N and S may be located on any interior position ofheterohydrocarbyl (include the positions on which the hydrocarbon groupattached to the remainder of the molecule). Embodiments comprise but arenot limited to —CH₂—CH₂O—CH₃, —CH₂—CH₂—NH—CH₃, —CH₂—CH₂—N(CH₃)—CH₃,—CH₂—S—CH₂—CH₃, —CH₂—CH₂, —S(O)—CH₃, —CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃,—CH₂—CH═N—OCH₃ and —CH═CH—N(CH₃)—CH₃. Up to two heteroatoms may beconsecutive, such as —CH₂—NH—OCH₃.

Unless otherwise specified, the terms “cycloalkyl”, “heterocycloalkyl”or a specific concept thereof (such as aryl, heteroaryl, cycloalkyl,heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl,heterocycloalkynyl, etc.) by itself or in combination with other termsrepresent, respectively, a cyclized “hydrocarbon group”,“heterohydrocarbyl”. In addition, for heterohydrocarbyl orheterocycloalkyl (such as heteroalkyl, heterocycloalkyl), the heteroatommay occupy the position at which the heterocycle is attached to the restof the molecule. Examples of cycloalkyl include, but are not limited to,cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl andthe like. Non-limiting examples of heterocyclic group include1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl,3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl,tetrahydrofuranindole-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl,1-piperazinyl and 2-piperazinyl.

The compounds of the present invention may be prepared by varioussynthetic methods well known to those skilled in the art, including thespecific embodiments enumerated below, embodiments in conjunction withother methods of chemical synthesis, and the equivalents familiar forthe skilled in the art. The preferred embodiments include but are notlimited to the examples of the present invention.

The solvents used in the present invention are commercially available,and can be used without further purification.

Compounds are named manually or with ChemDraw® software, and commercialcompounds are available under Supplier directory name.

The X-ray powder diffraction method is as follows:

Instrument: Bruker D8 ADVANCE X-ray diffractometer; Target: Cu: K-Alpha;Wavelength λ=1.54179{acute over (Å)}; Tube Voltage: 40 kV; Tube Current:40 mA; Scanning Range: 4-40; Sample Rotation Speed: 15 rpm; ScanningSpeed: 10°/min.

Unless otherwise specified, the following abbreviations are used herein:SGF represents simulated gastric fluid; Fassif represents simulatedintestinal fluid in fasting state; LDA represents lithiumdiisopropylamide; MsCl represents methanesulfonyl chloride; BOCrepresents t-butylcarbonyl as an amine protecting group; THF representstetrahydrofuran; rt represents room temperature; DCM representsdichloromethane; TEA represents triethylamine; DMF representsN,N-dimethylformamide; DBU represents 1,8-diazabicycloundec-7-ene; LAHrepresents lithium aluminum hydride; HATU represents2-(7-azobenzotriazole)-N,N,N′,N′-tetramethyluronium hexafluorophosphate;CDI represents carbonyldiimidazole; DMSO represents dimethylsulfoxide;MeOH represents methanol; EtOH represents ethanol; EtOAc representsethyl acetate; THF represents tetrahydrofuran; IPA representsisopropanol; ACN represents acetonitrile; MEK represents butanone; MIBKrepresents methyl isobutyl ketone; MTBE represents methyl tert-butylether; DPBS represents DuPont phosphate buffer solution; T3P representspropyl phosphoric anhydride; EDCI represents1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; HOBt represents1-hydroxybenzotriazole.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a XRPD pattern of Cu—K alpha radiation for crystal form I ofcompound 5-3.

FIG. 2 is a XRPD pattern of Cu—K alpha radiation for crystal form II ofcompound 5-3.

FIG. 3 is a XRPD pattern of Cu—K alpha radiation for crystal form III ofcompound 5-3.

FIG. 4 is a XRPD pattern of Cu—K alpha radiation for crystal form IV ofcompound 5-3.

DETAILED DESCRIPTION

The present invention will be further illustrated below with referenceto the specific examples, but these examples are not to make anydisadvantage limitation to the invention. This invention has beendescribed in detail herein, and the embodiments for carrying out theinvention are disclosed. It would be obvious for those skilled in theart can to make various changes and modifications to the embodiments forcarrying out the present invention without departing from the spirit andscope of the present invention.

Example 1: Preparation of Compound 1

Step 1: Preparation of Compound 3

22 liters of anhydrous toluene were added to a 50 liters reactor, sodiumhydrogen (2.2 Kg, 55.5 mol) was added slowly into the toluene solutionin batches under the protection of nitrogen, and compound 2 (3.93 Kg,33.3 mol) was dropped into the reactor at the same time. The compound 1(5.0 Kg, 22.2 mol) was dissolved in toluene (9 L) in a flask. Thereactor was heated to 85-95° C., and the toluene solution of compound 1prepared was added slowly to the reactor within about 2 hours. Payattention to the dripping rate and controlled the temperature at about95° C. After the addition was completed, the solution was stirred foranother 0.5-1 hour at this temperature, and then some samples were takenand quenched by water. The upper organic phase was checked by TLC andthe reaction was complete.

The reaction solution was cooled to about 60° C., and the viscousreaction solution was released while heat, and was poured into a mixedsolution of 66 liters of ice water, 32.5 liters of ethyl acetate and4.65 liters of hydrochloric acid to quench the reaction. Thecorresponding additional ice was supplemented to control temperaturebelow 10° C. during stirring. After the quench was completed, the layerwas statically separated, and the aqueous phase was extracted with 10liters of ethyl acetate, and the combined organic phase was washed with11 liters of saturated brine. The washed organic phase was dried over2.8 Kg of anhydrous sodium sulfate. After filtration, the productsolution was concentrated at 55° C. under reduced pressure (−0.09 Mpa)to give 7.25 Kg crude product which was used for the next step directlywithout further purification.

Step 2: Preparation of Compound 4

In a 50 liter reactor, 7.25 Kg of crude compound 3 was dissolved in 32liters of ethanol, and then cooled to 0-5° C. Sodium borohydride (843.6g, 22.2 mol) was added to the reactor in batches, and controlled thespeed of adding and made the temperature below 10° C., this took 0.5-1.5hours. After the addition was completed, stirring was continued at 0-10°C. for 2-3 hours. Samples were taken and monitored, TLC (ethylacetate/petroleum ether=1/3, high temperature coloration of ninhydrin)showed that the reaction was completed.

The temperature was controlled below 10° C., and the hydrochloric acid(11.1 L, 2M) was slowly dripped into the reactor to adjust pH to 6-7.After quenching, the reaction mixture was concentrated directly underreduced pressure (−0.09 Mpa, <55° C.) to remove most of the ethanol. Theconcentrated mixture solution was extracted with ethyl acetate two times(11 L×2), and the organic phase was combined and washed with saturatedbrine (5.5 L), dried over anhydrous sodium sulfate (2.2 Kg) andfiltered. The product solution was concentrated under reduced pressure(−0.09 Mpa, <55° C.) to give the oily crude product which was directlyused for the next step without purification.

Step 3: Preparation of Compound 5

In a 50 liter reactor (50 L), compound 4 (6.82 Kg, 22.2 mol, crude) wasdissolved in dichloromethane (30 L), and triethylamine (5.80 Kg, 57.3mol) was added. The temperature was cooled below 10° C., and themethanesulfonyl chloride (3.82 Kg, 33.3 mol) was slowly dripped into thereactor, kept the temperature below 15° C. for 1-2 hours. The reactionsolution was heated to 25° C. and stirred for 16 hours, samples weretaken. TLC (ethyl acetate/petroleum ether=1/3, high temperaturecoloration of ninhydrin) showed that compound 4 disappeared and thereaction was completed.

The reaction solution was washed with water (11 L) twice, and thenwashed with dilute hydrochloric acid (about 7 L, 2M). Controlled theaqueous phase pH=6-8, and then washed with saturated brine (11 Kg), anddried over anhydrous sodium sulfate (1.57 Kg) finally, filtered to givethe product solution, concentrated (−0.09 Mpa, <55° C.) to obtain thecrude product which was directly used for the next step withoutpurification.

Step 4: Preparation of Compound 6

In a 50 liter reactor, compound 5 (9.4 Kg, crude, 22.2 mol) wasdissolved in DMF (22 L), and the temperature was controlled below 40° C.DBU (6.76 Kg, 44.4 mol) was added to the reactor. After the addition wascompleted, the temperature was raised to 100° C. and stirred for 16-20hours. Samples were taken, TLC showed that compound 5 disappeared andthe reaction was completed.

The reaction solution was cooled to 25° C. Water (22 L) and ethylacetate (33 L) were then added and stirred for 10 minutes. The liquidwas layered, which the aqueous phase was extracted with ethyl acetate(22 L). The organic phase was combined, and washed with 0.5Nhydrochloric acid solution (15 L) (controlled the aqueous phase pH=6-7),the organic phase was washed with water (22 L) again. It was then driedover and filtered with anhydrous sodium sulfate (2.2 Kg). The solutionwas mixed with silica gel, and concentrated (−0.09 Mpa, <55 C) todryness to get the crude product which was purified by the column(eluent: ethyl acetate/n-heptane=1/20) to get the pure product compound6 (2.80 Kg, the total yield of four steps: 44.9%). ¹HNMR (CDCl₃, 400MHz) δ 6.69 (s, 1H), 4.75 (s, 1H), 4.30 (s, 1H), 4.16 (d, J=6.4 Hz, 2H),2.81 (bs, 1H), 2.12 (m, 1H), 2.02 (m, 1H), 1.92 (m, 1H), 1.62 (m, 1H),1.41 (s, 9H), 1.24 (t, J=7.2 Hz, 3H).

Step 5: Preparation of Compound 7-a and 7-b

Ethanol (1 L) was added to a 10 L autoclave, and the wet Pd/C (170 g)was slowly added into the autoclave for one time under the argonprotection. Compound 6 (563 g, 2.0 mol) was dissolved in ethanol (4.63L) and added to the autoclave. After the addition was completed, thereaction system was sealed, evacuated, and replaced with hydrogen twiceand then hydrogen was introduced. The initial pressure of hydrogen inthe reaction system was controlled under 4.0 Mpa, stirred overnight at10-15° C. Samples were taken, and the reaction was monitored by TLC orHPLC.

The reaction mixture was filtered through celite. The filter cake waswashed with ethanol (I L*2, 0.79 Kg*2). The filtrate was combined,concentrated under reduced pressure to give the product compounds 7-a,7-b (552 g, 7-a/7-b=3:1, yield: 97.5%). The product was used directlyfor the next step without purification.

Step 6: Preparation of Compound 8-a and 8-b

In a reactor (50 L), compound 7-a, 7-b (4.2 Kg, 7-a/7-b=3:1, 14.8 mol)was dissolved in anhydrous toluene (20 L), the reaction temperature wascontrolled between 0-5° C. The red aluminum (5.1 Kg, 17.8 mol) wasslowly added to the reactor under stirring, and the temperature was keptbetween 0-5° C. for about 1 hour. After the addition was completed,stirring was continued for 1 hour at 0-5° C. Samples were taken, TLC(ethyl acetate/petroleum ether=1/3, high temperature coloration) and GCmonitoring showed that the reaction was completed.

The temperature was controlled at 0-5° C. 15% sodium hydroxide solution(9 L), water (4.5 L) were added slowly to the reaction solution. Thetemperature was raised to 15° C. and stirred for 20 minutes. The aqueousphase of the system was extracted by toluene (10 L). The organic phasewas combined and washed with water (10 L), and then dried over anhydroussodium sulfate (1 Kg) finally. The product solution was obtained byfiltration and concentrated under reduced pressure to get compound 8-aand 8-b (3.525 Kg, yield: 98.7%). The compound was used directly for thenext step without purification. ¹HNMR (CDCl₃, 400 MHz) δ4.35 (m, 1H),4.13 (m, 1H), 3.47 (m, 1H), 2.05 (m, 1H), 1.82 (m, 1H), 1.73 (m, 2H),1.67 (m, 3H), 1.45 (s, 9H), 1.28 (m, 2H).

Step 7: Preparation of Compound 10-a and 10-b

In a 50 L reactor, compound 8-a and 8-b (3.425 Kg) were dissolved intetrahydrofuran (17 L), and sodium hydroxide (1.136 Kg) and compound 9(1.960 Kg) were added to the reactor. Controlled the temperature under75° C. and stirred for 24 hours. Samples were taken, GC and TLC (ethylacetate/petroleum ether=1/3, high temperature coloration ofphosphomolybdic acid) detected that the raw materials reactedcompletely.

The reaction solution was cooled to room temperature, and the saturatedbrine (8.5 L) was added into the solution. Stirring was continued for 5minutes. The mixture was separated. The organic phase was adjusted to pH6-7 with 2.0 N hydrochloric acid. The mixture was separated. The aqueousphase was combined, and adjusted to pH 6-7 with 2.0 N hydrochloric acid,and extracted with ethyl acetate (10 L). The organic phase was combinedand dried over anhydrous sodium sulfate (1 Kg) and filtered. The productsolution was concentrated under reduced pressure (−0.09 Mpa, <55° C.) toget the crude product which was purified by silica gel columnchromatography (n-heptane:ethyl acetate=20:1 to 5:1) to get a lightyellow oily product (4.5 Kg yield 92.9%). ¹HNMR (CDCl₃, 400 MHz) δ7.95(s, 1H), 7.32 (m, 1H), 6.69 (m, 1H), 4.38 (m, 2.5H), 4.13 (m, 1.5H),2.03 (m, 3H), 1.86 (m, 3H), 1.45 (m, 3H), 1.26 (s, 9H).

Step 8: Preparation of Compound 16

In a reactor, compound 15 (2.5 Kg, 9.54 mol) was dissolved in anhydrousmethanol (10 L), and the concentrated sulfuric acid (360 mL, 7.16 mol)was slowly dripped into the reactor at room temperature. The reactor washeated to 65-80° C., and reacted for 18 hours. HPLC monitored that 5-7%of raw materials still remained. The reaction was supplemented withconcentrated sulfuric acid (36 mL, 67.3 g) and continued to react for 18hours. HPLC detected that the raw materials reacted completely.

The reaction solution was cooled to 26° C. (four-pot reactions mergedfor treatment), and 10N sodium hydroxide solution (3.75 L) was slowlyadded to adjust pH to 5-6. The reaction solution was concentrated toabout 35 L, and ethyl acetate (20 L), water (8 L), and 7% of sodiumbicarbonate (10 L) were added into the reaction solution. Stirred andseparated, the aqueous phase was extracted again with ethyl acetate (20L), the organic phase was combined and washed with saturated brine (15L), and dried over anhydrous sodium sulfate. The product solution wasconcentrated (−0.09 MPa, <55° C.) to obtain 2.5 Kg of product (yield:95%) which was directly used for the next step without purification.¹HNMR (DMSO-d6, 400 MHz) δ7.76 (d, J=8.4 Hz, 1H), 7.45 (s, 1H), 6.98 (d,J=8.0, 2.5H), 3.77 (s, 3H), 2.19 (s, 3H).

Step 9: Preparation of Compound 18

First, compound 16 (5 Kg, 18.1 mol), compound 17 (4.83 Kg, 19.0 mol) andpotassium acetate (4.44 Kg, 45.3 mol) were added to a reactor, and thenanhydrous DMF (20 L) was added to the reactor. Stirred and dissolved atroom temperature. Palladium acetate (122 g, 54.33 mmol) was added underthe protection of nitrogen, the reactor was heated to 80-90° C., thereaction was stirred for 16 hours. Samples were taken, TLC (ethylacetate/petroleum ether=1/10, UV 254 nm) and HPLC detected that thematerial 16 reacted completely.

The reaction solution was cooled to room temperature (25-34° C.), thenfiltered through celite, and washed with ethyl acetate (post-processingof two-pot reactions merged at this step). The filtrate was added ethylacetate and water, stirred and separated. The organic phase was washedwith sodium bicarbonate solution (5 L, 8%) and saturated brine (5 L)respectively, dried over anhydrous sodium sulfate (3 Kg) finally,filtered, and the product solution was concentrated to get 8 Kg of crudeproduct (yield: 80%) which was used directly for the next step withoutpurification.

Step 10: Preparation of Compound 20

First, compounds 18 (4 Kg, crude, 14.485 mol), compound 19 (1.74 Kg,15.2 mol), and sodium carbonate (3.5 Kg, 33 mol) were added to a 50 Lreactor, and then 2-methyl tetrahydrofuran (25 L) and water (8 L) wereadded to the reactor, stirred and dissolved at room temperature. Pd(dppf) Cl₂.DCM (325 g, 0.398 mol) was added under the protection ofnitrogen, the reactor was heated to 70-80° C., and the reaction wasstirred for 16 hours. Samples were taken for monitoring: TIC (ethylacetate/petroleum ether=1/2) showed that the raw materials reactedcompletely.

The reaction solution was cooled to room temperature (30° C.), water (10L) was added and stirred for 10 minutes. The mixture was separated. Theaqueous phase was filtered through celite and extracted with ethylacetate (10 L). The organic phase was combined, activated carbon powder(2.3 Kg) was added, and stirred for 2 hours (two-pot reactions merged atthis step). The filtrate was filtered through celite and concentrated(<50° C.) until solid precipitation, filtered to get a black solid. Thesolid was dissolved in ethyl acetate (3-5 L) and filtered by silica gelcolumn, washed with ethyl acetate, and concentrated to get compound 20(3.0 Kg, yield: 45.5%).

Palladium Removal Process:

Compound 20 (3.0 Kg) was added to a reactor, dichloromethane (12 L) wasadded, stirred and dissolved at room temperature (25-27° C.). Sodiumchloride (210 g), thiocyanuric acid (27 g), ammonium hydroxide (225 mL),water (2500 mL) and ethanol (to 2400 mL) were formulated into TMTsolution (−5 L). The TMT solution (−5 L) was added to the reactor, andthe reaction mixture was stirred at room temperature (20-25° C.) for 24hours. The reaction solution was filtered through celite, the aqueousphase was separated and removed, and the organic phase was concentratedto dryness (−0.09 Mpa, <50° C.) to get the compound 20 (2.92 Kg, yield:97.3%). ¹HNMR (DMSO-d6, 400 MHz) δ8.87 (d, J=4.8 Hz, 2H), 7.96 (d, J=8.0Hz, 1H), 7.46 (m, 3H), 3.62 (s, 3H), 2.41 (s, 3H).

Step 11: Preparation of Compound 14

In a 50 L reactor, compound 20 (2900 g, 12.7 mol) was dissolved in2-methyl tetrahydrofuran (15 L). The sodium hydroxide solution (NaOH:1270 g, 31.8 mol; water: 20 L, 20 Kg) was prepared, and added to thereactor, the reaction solution was heated to 70-75° C. and stirred for16 hours, TLC (ethyl acetate/petroleum ether=1/2, UV: 254 nm) detectionshowed that the reaction finished.

Cooled, and the aqueous phase was separated and washed with ethylacetate (10 L). The temperature of aqueous phase was controlled below20° C., 12N hydrochloric acid (3.3 L) was slowly added to adjust pHto 1. The temperature was kept less than 30° C. In this process, thesolid was precipitated, stirred for about 10 minutes, filtered, thefilter cake was rinsed with water (5 L*2). The filtered solid was driedin vacuum to get the product compound 14 (2600 g, purity: 99%, yield:95.6%). ¹HNMR (DMSO-d6, 400 MHz) δ12.67 (br, s, 1H), 8.85 (d, J=4.0 Hz,2H), 7.81 (d, J=7.6 Hz, 1H), 7.48 (s, 1H), 7.43 (m, 2H), 2.40 (s, 3H).

Step 12: Preparation of Compound 12-a and 12-b

In a reactor (30 L), compounds 10-a and 10-b were dissolved in ethylacetate (13.5 L). The temperature was controlled at 10-20° C., and 35%of concentrated hydrochloric acid (4.5 L) was added to the reactor. Thetemperature was controlled less than 15° C. (paid attention to controldrip rate). After the addition was complete, the temperature wascontrolled at 20-25° C. and reacted for 16-20 hours, samples were takenfor control. The HPLC test showed that the reaction was completelyfinished.

Water (5 L) was added and layered, the water layer was collected. Theorganic layer was extracted again with water (5 L). The water layer wascombined, washed with ethyl acetate (5 L); the water layer was separatedand cooled to below 10° C. The pH of water layer was adjusted to 14 withsodium hydroxide (3.0 Kg) (the addition speed of base was noted,controlled the inner temperature below 20° C.), the water layer wasextracted with ethyl acetate (10 L+5 L) twice. The organic layer (15.5Kg) was collected and the samples were taken for test. Purity=82.8%,Content=16.4%, the product weight was calculated: 2.54 Kg. The ethylacetate solution containing the product was used directly for the nextstep without other treatment.

Step 13: Preparation of Compound 13

A solution of compound 12-a and compound 12-b (15.5 Kg, containing 2.54Kg of compound 10) was added to the reactor, and heated to 55-65° C. Asolution formulated by D-tartaric acid (1.62 Kg) and water (3.5 L) wasslowly dripped into the reactor. After the addition was complete, thereaction was continued at 55-65° C. for 2-3 hours. Slowly cooled to5-15′C and stirred overnight, filtered to obtain the wet product (5.78Kg). The wet product and water (15 L) was added to the reactor, andheated to 80-90° C. to dissolve clearly, then slowly cooled to 15-25° C.and stirred continuously for 16-20 hours. Filtered to get the wetproduct (2.6 kg), samples were taken for test, e.e.=90.8%. The wetproduct and water (6 L) was added to the reactor, and heated to 80-90°C. to dissolve clearly, slowly cooled to 15-25° C. within 16-20 hours.Filtered to get the wet product (2.2 kg), samples were taken for test,e.e.=99.6%.

The wet product (2.2 Kg) and H₂O (2 L) were added to the reactor andstirred to dissolve. The formulated K₂CO₃ aqueous solution (K₂CO₃:3.55Kg, water: 10 L) was added slowly to the reactor, and the temperaturewas controlled at 15-25° C., stirred for 0.5-1 hours. The reactionsolution was extracted with ethyl acetate (10 L*2), the organic phasewas combined, dried over 2.0 Kg anhydrous Na₂SO₄ and filtered. Thefiltrate was concentrated to dryness (−0.09 MPa, <50° C.) to obtain acolorless oil compound 13 (1006 g) which turned into a white solid aftercooling. Samples were taken for test ee=99.8%, HPLC Purity: 99.7%. Theproduct was used directly for the next step without further treatment.

Step 14: Preparation of Compound 5-3

In a reactor, compound 13 (630 g, 2.67 mol) was dissolved in ethylacetate (7.9 L), and compound 14 (685 g, 3.20 mol) was added to thereactor and stirred for 20 minutes, the temperature was controlled at15-25° C., T₃P (1903 mL, 3.20 mol) was slowly added, after 20-30 minutesof addition, the reaction solution was heated to 40-60° C. and stirredfor 2-3 hours. Samples were taken for monitoring. HPLC showed that theratio of raw material to product was less than 2.5%.

Cooled to 15-25° C. and H₂O (7.9 L) was added, stirred for 0.5-1 hours,the organic phase was separated, and the aqueous phase was extractedwith ethyl acetate (7.9 L). The organic phase was combined and washedwith water (7.9 L) for one time. The solvent was evaporated underreduced pressure (−0.09 MPa, <50° C.) to get the crude compound 15 (1.3KG). The crude compound was added to 3.0 L of ethyl acetate, slurried atroom temperature and filtered to get the target compound (1.1 Kg, yield:95.4%). HPLC Purity: 99.9%, e.e value: 100%. XRPD test showed it wascrystal form I. ¹H NMR (400 MHz, DMSO-d6)=8.83 (br. s., 2H), 8.17-8.01(m, 2H), 7.49-7.33 (m, 3H), 6.86 (dd, J=3.5, 9.0 Hz, 1H), 6.86 (dd,J=3.5, 9.0 Hz, 1H), 4.63 (br. s., 1H), 4.43 (m, 1H), 4.11 (br. s, 1H),3.79 (m, 1H), 2.52-2.48 (m, 2H), 2.35-1.95 (m, 8H), 1.29-1.20 (m, 2H).

Step 15: Preparation of Crystal Form II

Crystal form I (1.1 Kg), ethyl acetate (2750 mL) and isopropanol (110mL) were added into a 51 reaction flask, the reaction system was heatedto 65-80° C. and stirred for 2-3 hours, the temperature was slowlylowered to 50-60° C. within 1-2 hours, the suspension was continuouslystirred at 40-50° C. for 20-24 hours. A little of samples were taken,and detected by XRPD which showed the crystal completely transformedsuccessfully. The temperature of system was cooled to 10-20° C. slowly,and kept stirring at this temperature for 20-24 hours. The XRPD testshowed that the crystal form was not changed. A solid was obtained byfiltration and dried at 40-50° C. in vacuum for 20-24 hours to get theproduct (660 g, yield: 60%). HPLC Purity: 99.13%, ee value: 100%. TheXRPD test showed it was crystal form II.

Example 2: Preparation of Crystal Form III

0.30 g of crystal form 1 of compound 5-3 was added to 4.5 ml toluene,fully dissolved and filtered, 4 ml n-heptane was gradually added to thefiltrate under stirring until the solid was precipitated, continuouslystirred at 20-30° C. for 16-22 hours, filtered to obtain a solid, driedunder vacuum at 40° C. to get 0.22 g of crystal form III (yield: 73.3%).

Example 3: Preparation of Crystal Form IV

0.30 g of crystal form I of compound 5-3 was added to 4.5 ml ethanol,fully dissolved and filtered, 9 ml pure water was gradually added to thefiltrate under stirring until the solid was precipitated, continuouslystirred at 20-30° C. for 16-22 hours, filtered to obtain a solid, driedunder vacuum at 40° C. to get 0.23 g of crystal form IV (yield: 76.7%).

Experimental Example 1: In Vitro Test of OX1/2R Experimental Purpose

The inhibitive effect of a compound on OX1 and OX2 GPCR receptor wasevaluated by detecting calcium signal change in cells with FLIPR, andIC50 value of compound was used as an indication.

Experimental Materials

-   -   Cell line: HEK293-OX1 and OX2 stable cell strain    -   HEK293-OX1 cell culture media (DMEM, Invitrogen #11960-044, 10%        serum Gibco #10099141, L-Glutamine 1×, Gibco #25030, sodium        pyruvate 1×, Gibco #11360, Geneticin 300 μg/ml, Gibco #10131).    -   HEK293-OX2 cell culture media (DMEM, Invitrogen #11960-044, 10%        serum Gibco #10099141, L-Glutamine 1×, Gibco #25030, sodium        pyruvate 1×, Gibco #11360, Geneticin 300 μg/ml,    -   Gibco #10131, Blasticin 2 μg/ml, Invitrogen #R21001).    -   Pancreatic enzyme (Invitrogen, #25200-072)    -   DPBS (Hyclone, #SH30028.01B)    -   Fluo-4 AM, Invitrogen #F14202    -   F-127, Invitrogen #P3000MP    -   Probenecid, Sigma #P8761    -   384-well cell plate, Greiner #781946    -   384-well compound plate, Greiner #781280    -   CO₂ incubator, Thermo #371    -   Centrifuge, Eppendorf #5810R    -   Vi-cell cytometry, Beckman Coulter    -   POD 810 Plate Assembler Automatic microplate pretreatment system    -   Labcyte FLIPR, Molecular Device.

Experimental Procedures and Methods

a) Cell inoculation (HEK293-OX1 and HEK293-OX2 cells)

-   -   1) The medium, trypsin, and DPBS were preheated at 37° C. under        water bath. Culture medium of cells was sucked and cells were        washed with 10 mLD PBS;    -   2) The preheated trypsin was added into the culture bottle which        was rotated so that trypsin uniformly covered the bottle. It was        placed in an incubator (37° C., 5% CO₂) to digest for 1-2        minutes;    -   3) Each T150 was suspended with 10-15 mL of culture medium, and        centrifuged at 800 rpm for 5 minutes. Cells were resuspended        with 10 mL medium, and 1 mL of the cell re-suspension was sucked        out and counted with Vi-cell cytometry.    -   4) The OX1 cells were diluted with culture medium to 5×10⁵        cells/mL, and OX2 cells were diluted to 4×10⁵ cells/mL. The        diluted cells were added into 384 plate (Greiner, 781946) with        multichannel pipettes (50 L/hole, OX1 cells: 25000 cells/hole;        and OX2 cells: 20000 cells/hole).

The cell plate was placed in an incubator (37° C., 5% CO₂) overnight.

b) loading of the compound:

-   -   1) DMSO was used to dilute the compound into 20 mM by using        3-fold dilution. 8 gradients in duplicate wells were used. Echo        liquid handler was used to add the compound into a compound        plate. Then 20 μL buffer was added to ensure that the final DMSO        concentration was 0.1%.        c) FLIPR experiment:

1) The cell culture medium in 384-well plate was washed away with avacuum pump. 30 μL fluorescent dye Fluo4AM was added. The cell wasincubated at 37° C., 5% CO₂ in an incubator for 1 hr and thenre-equilibrated under room temperature for 10 minutes.

2) EC50 Test: Orexin A was diluted manually on ice by using 3-folddiluted. 8 gradients in duplicate wells were used. Then the DMSO platewas prepared and the DMSO concentration was 0.5%. The cell plate, OrexinA plate, and DMSO plate were placed into FLIPR respectively, and thefluorescence values were read.

3) EC70 value was calculated based on EC50 value of Orexin A. 5×EC70solution was prepared and added into a 384-well compound plate withmultichannel pipettes. The plate was placed on ice for preservation.

4) In the FLIPR, the compound plate, 5×EC70 plate, cell plate and FLIPRtips were placed respectively. The program was run and the fluorescencevalues were read.

d) Data Analysis: Prism5.0 was used to analysis the data, and the IC₅₀value of the compound was calculated.

The experimental results are shown in table 1:

TABLE 1 IC₅₀ experimental results detected in FLIPR Test sample hOX1RhOX2R Compound 5-3 A A Note: A ≤ 50 nM.Conclusion: the compound of the invention show significant inhibitoryeffects on OX1 and OX2 GPCR receptors.

Experimental Example 2: Solubility Test

Solubility Study of crystal form I of compound 5-3:1-1.5 mg of crystalform I of compound 5-3 was added to a flask at room temperature, asingle organic solvent or a mixed solvent was added in small quantitiesand many times until the visual solution was clear or no solid particleswere present, determined the solubility of crystal form I in differentsolvents, the results were shown in Table 2.

TABLE 2 The solubility of crystal form I in different solventsSolubility Num. Solvent (mg/mL) 1 MeOH >53 2 EtOH 22-55 3 IPA 10-20 4n-butanol 22-55 5 ACN 20-50 6 Acetone >52 7 MEK >54 8 MIBK 24-60 9 EtOAc19-48 10 MTBE  <1 11 THF >53 12 Toluene >54 13 1,4-dioxane >57 14 water <1 15 MeOH—H₂O (3:1) 18-46

Solubility Study of crystal form II of compound 5-3:10 mg of crystalform II was added to a 1.5 mL volumetric flask at 37° C., 1 mL solventwas added, shook for 24 hours, centrifuged, and the supernatant wasimmediately analyzed by HPLC, the results were shown in Table 3,

TABLE 3 The solubility of crystal form II in different aqueous mediaAppear- Solubility Final Crystal form of Solvent ance (mg/mL) pHSolubility residual solid 0.1N HCl turbid 0.42 0.80 Slightly solublecrystal form II water turbid 0.14 7.95 Slightly soluble crystal form IISimulate turbid 0.27 1.88 Slightly soluble crystal form II gastric fluidSimulated turbid 0.18 6.65 Slightly soluble crystal form II fastingintestinal fluid Simulated turbid 0.29 5.04 Slightly soluble crystalform II postpran- dial intestinal fluid

What we claim:
 1. A crystal of compound 5-3 as shown in formula 5-3,

wherein the crystal is crystal form IV; the XRPD pattern analysis ofcrystal form IV is as follows: diffraction angle 2θ relative intensity(%) 8.653 100.0 10.408 94.7 12.106 45.5 12.362 15.5 13.723 14.5 14.1526.1 14.414 7.3 14.925 10.6 15.988 18.9 16.682 20.1 16.937 33.6 17.31179.1 17.982 61.7 18.338 29.2 19.245 5.9 19.856 29.2 20.721 24.1 21.0208.2 21.611 34.8 23.385 22.1 23.997 44.0 24.430 30.5 24.965 15.2 26.2259.3 26.562 23.9 26.977 6.8 27.647 16.3 27.983 10.9 29.659 3.1 30.842 3.031.788 12.4 33.031 2.7 33.426 2.2 33.858 2.8 34.884 2.3 36.499 4.837.427 2.9.


2. The crystal form of compound 5-3 of claim 1, wherein the crystal formis crystal form IV whose XRPD pattern is shown in FIG.
 4. 3. A processfor preparing the crystal form of compound 5-3 of claim 1, comprising:adding crystal form I of compound 5-3 into a second organic solvent,dissolving at room temperature or heating to a temperature of 40° C. toreflux temperature for dissolution, then slowly adding a third organicsolvent dropwise, and after the addition is completed, forming thecrystal form IV at room temperature or by cooling to 20-30° C. in 1-120hours; the second organic solvent is selected from methanol, ethanol,isopropanol, n-butanol, ethyl formate, ethyl acetate, isopropyl acetate,isobutyl acetate, n-butyl acetate, acetonitrile, dichloromethane and amixture thereof; the third organic solvent is water; and the XRPDpattern analysis of crystal form I of compound 5-3 is as follows:diffraction angle 2θ relative intensity (%) 8.611 100.0 10.330 70.612.284 13.4 13.703 23.9 14.057 5.6 14.351 4.8 14.848 9.2 15.576 7.516.602 31.6 17.250 71.3 17.902 90.7 18.277 42.4 19.149 6.5 20.625 21.721.550 42.7 22.124 2.3 23.307 33.4 23.953 2.6 24.370 32.3 26.501 29.426.915 8.7 27.663 11.0 27.939 15.5 29.499 2.6 30.604 2.2 31.727 13.332.969 2.6 33.378 2.8 33.777 3.0 34.825 1.6 36.399 5.3 37.366 4.3 39.3573.3.


4. The process according to claim 3, wherein the crystal form I isprepared by: adding compound 5-3 into ethyl acetate and pulping toprovide the crystal form I.
 5. The process of claim 4, wherein thecompound 5-3 is prepared by the following step:

wherein the condensation agent is selected from propyl phosphoricanhydride, carbonyldiimidazole,1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, 1-hydroxybenzotriazole,or 2-(7-azobenzotriazole)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate.
 6. The process of claim 5, wherein the process forpreparing compound 13 as shown in formula 13 comprises the followingstep:

wherein, the separation agent is D-tartaric acid; the reaction solventis selected from the group consisting of ethyl acetate, isopropylacetate, ethanol, methanol, tetrahydrofuran, dichloromethane, and water;the recrystallization solvent is selected from the group consisting ofwater and ethanol; and the base is selected from the group consisting ofsodium carbonate, potassium carbonate, sodium hydroxide, and potassiumhydroxide.